Human Cytochrome b-245 heavy chain (CYBB) ELISA Kit

Cytochrome b (-245) is composed of cytochrome b alpha (CYBA) and beta (CYBB) chain. It has been proposed as a primary component of the microbicidal oxidase system of phagocytes. CYBB deficiency is one of five described biochemical defects associated with chronic granulomatous disease (CGD) . In this disorder, there is decreased activity of phagocyte NADPH oxidase; neutrophils are able to phagocytize bacteria but cannot kill them in the phagocytic vacuoles. The cause of the killing defect is an inability to increase the cell's respiration and consequent failure to deliver activated oxygen into the phagocytic vacuole.

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Human Cytochrome b-245 heavy chain (CYBB) ELISA Kit has high sensitivity and excellent specificity for detection of Human CYBB. No significant cross-reactivity or interference between Human CYBB and analogues was observed.

This Human Cytochrome b-245 heavy chain (CYBB) ELISA Kit employs a two-site sandwich ELISA to quantitate CYBB in samples. An antibody specific for CYBB has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and anyCYBB present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for CYBB is added to the wells. After washing, Streptavidin conjugated Horseradish Peroxidase (HRP) is added to the wells. Following a wash to remove any unbound avidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of CYBB bound in the initial step. The color development is stopped and the intensity of the color is measured.

• Do not mix components from different kit lots or use reagents beyond the kit expiration date.• Allow all reagents to warm to room temperature for at least 30 minutes before opening.• Pre-rinse the pipet tip with reagent, use fresh pipet tips for each sample, standard and reagent to avoid contamination.• Unused wells must be kept desiccated at 4 °C in the sealed bag provided.• Mix Thoroughly is very important for the result. It is recommended using low frequency oscillator or slight hand shaking every 10 minutes.• It is recommended that all samples and standards be assayed in duplicate or triplicate.